FIGURE 5.

DSB repair modification by MS2-CtIP and SunL-CtIP fusion proteins in HEK^TLR6 reporter cells. (A) Expression vectors for Cas9 or Cas9-SunTag, sgRosa26(MS2), pTLR-donor and CtIP, MS2-CtIP, MS2^di-CtIP, or SunL-CtIP were cotransfected into HEK^TLR6 cells and the frequency of Venus⁺ cells (green columns) and RFP⁺ cells (red columns), indicating DSB repair by HDR or NHEJ, was determined by flow cytometry (FACS) 72 h after transfection. (B) Transfection results are shown as bar graphs representing mean values with standard deviation of triplicate samples. Plasmids selected (+) for individual transfection samples are indicated in the table below. Mean values were used to calculate the ratio of Venus⁺ to RFP⁺ cells and p-values (T-test) to determine the significance in levels of Venus⁺ or RFP⁺ cells between samples 2 and 3–6 (table bottom) and to compare samples 4 and 5 (top horizontal lines). n.s., not significant, p > 0.05. Two independent replicates of the assay were performed that confirmed the results.